State Key Laboratory of Applied Microbiology Southern China, Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, Guangdong Institute of Microbiology, Guangdong Academy of Sciences, Guangzhou, 510070, China.
Sunnybrook Research Institute, Sunnybrook Health Sciences Centre, Toronto, M4N3M5, Canada.
Int J Biol Sci. 2020 Jan 1;16(1):61-73. doi: 10.7150/ijbs.38475. eCollection 2020.
has long been used as the health and medicinal plant known to induce cancer cell death. However, the bioactive compounds of and the underlying mechanism of their regulator in cell autophagy and cell apoptosis remain unexplored. In our previous study, we found that the ethanol extract had antitumor activity through inducing cancer cell death. In this study, by detecting the anti-tumor effect of sequence extracts from , the active constituents were collected in solvent ethyl acetate. A strategy based on ultra-performance liquid chromatography coupled with hybrid quadrupole-orbitrap mass spectrometry (UPLC-Q-Orbitrap/MS) was first utilized to analyze the chemical constituents of active fraction (ethyl acetate fraction, CS3). A total of 29 compounds including 8 triterpenoids, 6 flavonoids, 4 fatty acids, 8 phenolic acids, 1 anthraquinones, 1 nucleoside and 1 sterol were detected and identified or tentatively identified for the first time in . We found that CS3 induces cancer cell death accompanied with a great number of vacuoles in the cytoplasm. CS3-induced autophagosome formation was found and confirmed by electron microscopy and the high expression levels of microtubule-associated protein-1 light chain 3-II (LC3II), a marker protein of autophagy. We additionally demonstrated that CS3 activated and increased the pro-apoptotic mitochondrial proteins, BNIP3 and BNIP3L, in mRNA and protein levels. The constituents of CS3 down-regulated anti-apoptotic BCL2, and then releases autophagic protein Beclin-1. These finding for the first time systematically not only explore and identify the active constituents of CS3 in , but also examined the mechanism associated with CS3-induced cell death via cell autophagy. This active component may serve as a potential source to obtain new autophagy inducer and anti-cancer compounds for hepatocellular carcinoma.
已长期被用作具有诱导癌细胞死亡作用的保健和药用植物。然而，[植物名称]的生物活性化合物及其在细胞自噬和细胞凋亡中的调节机制仍未得到探索。在我们之前的研究中，我们发现乙醇提取物通过诱导癌细胞死亡具有抗肿瘤活性。在这项研究中，通过检测[植物名称]系列提取物的抗肿瘤作用，用溶剂乙酸乙酯收集活性成分。首先利用基于超高效液相色谱与混合四极杆轨道阱质谱（UPLC-Q-Orbitrap/MS）的策略分析活性部位（乙酸乙酯部位 CS3）的化学成分。共检测并鉴定了 29 种化合物，包括 8 种三萜、6 种黄酮、4 种脂肪酸、8 种酚酸、1 种蒽醌、1 种核苷和 1 种甾体，这在[植物名称]中是首次发现。我们发现 CS3 诱导癌细胞死亡，同时细胞质中出现大量空泡。通过电子显微镜观察到 CS3 诱导自噬体形成，并得到证实，微管相关蛋白 1 轻链 3-II（LC3II）是自噬的标志物蛋白，其表达水平升高。此外，我们还证明 CS3 激活并增加了促凋亡的线粒体蛋白 BNIP3 和 BNIP3L 的 mRNA 和蛋白水平。CS3 的成分下调了抗凋亡的 BCL2，然后释放自噬蛋白 Beclin-1。这些发现首次系统地探索和鉴定了 CS3 在[植物名称]中的活性成分，同时研究了 CS3 诱导细胞死亡与细胞自噬相关的机制。该活性成分可能成为获得新的自噬诱导剂和肝癌抗癌化合物的潜在来源。
